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Analysis of the transcriptome of differentiating and non-differentiating preadipocytes from rats and humans by next generation sequencing

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Abstract

Alongside cell lines such as 3T3-L1 cells, primary cell culture models of adipogenesis have helped in developing an understanding of the process of adipocyte recruitment and maintenance, which may lead to therapeutic advances to treat the growing epidemic of obesity. Recently, it has been demonstrated that fat cell progenitors (DFAT) established through ceiling culture of adipocytes retain an enhanced ability to undergo adipocyte differentiation compared to preadipocytes isolated from the stromal vascular fraction of adipose tissue. Clonal expansion of rat DFAT cells identified differentiation capable and incapable cell strains. To understand the mechanisms underlying these differences, comparison of their transcriptomes by next generation sequencing was performed. Two hundred seventy-eight genes with a significant fold change of 1.4 were detected as being consistently deregulated between differentiating and non-differentiating strains. Bioinformatic network analyses identified components of the extra-cellular matrix and PPARγ as important genes in this process, suggesting crosstalk between ECM and transcription factors influences differentiation. Analyses of the transcriptomes of human DFAT cells in early and late passage (non-differentiating) confirmed the importance of these pathways in maintaining an adipogenic potential.

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Acknowledgments

We would like to thank Werner Rust, Heike Günther, Silke Rist, and Sigrid Schacherl-Schmid for their excellent technical assistance.

Conflict of interest

All the authors were employed by Boehringer Ingelheim Pharma GmbH & Co. KG during the time the study was conducted.

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Authors and Affiliations

  1. Target Discovery Research, Boehringer Ingelheim Pharma GmbH & Co. KG, Birkendorferstraße 67, 88397, Biberach an der Riß, Germany

    Fabian Birzele & Tobias Hildebrandt

  2. CardioMetabolic Diseases Research, Boehringer Ingelheim Pharma GmbH & Co. KG, Birkendorferstraße 67, 88397, Biberach an der Riß, Germany

    Sybille Fässler, Heike Neubauer & Bradford S. Hamilton

Authors
  1. Fabian Birzele
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  2. Sybille Fässler
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  3. Heike Neubauer
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  4. Tobias Hildebrandt
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  5. Bradford S. Hamilton
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Corresponding author

Correspondence to Bradford S. Hamilton.

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Birzele, F., Fässler, S., Neubauer, H. et al. Analysis of the transcriptome of differentiating and non-differentiating preadipocytes from rats and humans by next generation sequencing. Mol Cell Biochem 369, 175–181 (2012). https://doi.org/10.1007/s11010-012-1380-1

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  • DOI: https://doi.org/10.1007/s11010-012-1380-1

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